Composition containing cross-linked collagen, preparation method and application of composition in wound repair

Abstract

The invention belongs to the technical field of cosmetics, and particularly relates to a composition containing cross-linked collagen, a preparation method of the composition and application of the composition in the aspect of wound repair. The composition is prepared from the following components in percentage by mass: 0.1 to 0.3 percent of mild crosslinking type I collagen, 0.05 to 0.1 percent of type III collagen, 0.1 to 0.2 percent of mussel mucin, 1 to 3 percent of humectant, 1.5 to 3 percent of preservative and the balance of water. According to the preparation method disclosed by the invention, a heterogeneous fiber network is constructed by lightly crosslinked type I collagen and type III collagen prepared by a specific process according to a specific proportion through a step-by-step assembly technology for the first time, and mussel mucin is introduced as a dynamic crosslinking agent; the three-dimensional hydrogel composition with a triple structure of I type collagen skeleton-III type collagen soft network-mussel mucin crosslinking point is formed through self-assembly under physiological conditions, and the composition can provide a more excellent bionic microenvironment for tissue repair, so that the wound repair effect is remarkably improved.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, and in particular relates to a composition containing cross-linked collagen, its preparation method, and its application in wound repair. Background Technology

[0002] Wound repair and tissue regeneration are major challenges that continue to be a focus of attention in the biomedical field. Ideal wound repair materials should be able to mimic the structure and function of the natural extracellular matrix, providing a suitable three-dimensional scaffold for cell adhesion, migration, and proliferation, while maintaining a moist healing environment and possessing sufficient mechanical strength to protect the wound.

[0003] Collagen, as a major component of the extracellular matrix, is widely considered the gold standard for wound repair materials due to its excellent biocompatibility, biodegradability, and ability to be recognized and utilized by cells. Type I collagen provides the main mechanical strength of tissues, while type III collagen is closely related to early embryonic development and scarless healing; both coexist in a certain proportion in young, healthy skin. However, single-collagen-based materials have significant limitations in practical applications, such as insufficient mechanical strength, rapid degradation, poor adhesion to moist wounds, and limited functionality and synergistic effects, which severely restrict their repair efficacy and clinical application value.

[0004] Therefore, there is an urgent need in this field to develop a novel wound repair composition that can simultaneously address the core pain points of insufficient mechanical strength, poor adhesion, and limited and poor functionality, in order to break through the existing technological bottlenecks. Summary of the Invention

[0005] The purpose of this invention is to overcome the shortcomings of the prior art and provide a composition containing cross-linked collagen, a preparation method thereof, and its application in wound repair.

[0006] To achieve the above objectives, the technical solution adopted by the present invention includes: In a first aspect, the present invention provides a composition containing cross-linked collagen, the composition comprising the following components in weight percentages: 0.1-0.3% mildly cross-linked type I collagen, 0.05-0.1% type III collagen, 0.1-0.2% mussel adhesive protein, 1-3% humectant, 1.5-3% preservative, and the balance being water.

[0007] Preferably, the mass ratio of the mildly cross-linked type I collagen to type III collagen is (2-3):1.

[0008] More preferably, the mass ratio of the mildly cross-linked type I collagen to type III collagen is 3:1.

[0009] This invention breaks through the limitations of single collagen or simple mixture systems, and for the first time organically combines mildly cross-linked type I collagen, type III collagen, and mussel adhesive protein to form a functionally complementary system of "skeleton-soft network-dynamic cross-linking agent". Each component has a clearly defined and irreplaceable function, addressing the three core issues of mechanical support, cell guidance, and wound adhesion, thus avoiding the shortcomings of traditional compositions that suffer from single function or insufficient synergy. Furthermore, this invention specifies a particular ratio between mildly cross-linked type I collagen and type III collagen to simulate the structure of the natural extracellular matrix, thereby constructing a heterogeneous fiber network with "type I collagen as the backbone and type III collagen as branches". This avoids insufficient repair efficiency or structural instability caused by an excessively high proportion of a single collagen, and provides cells with a growth matrix that most closely resembles the physiological environment.

[0010] Based on the specific raw material selection and optimized formulation of this invention, the resulting three-dimensional network hydrogel possesses four major characteristics: strength (slightly cross-linked type I collagen), flexibility (type III collagen), adhesion (mussel adhesive protein), and maintenance of a moist environment. This makes it suitable for irregular wound surfaces with exudate. Furthermore, early-stage type III collagen guides cell migration and proliferation, while later-stage lightly cross-linked type I collagen provides long-term mechanical support, preventing scar contracture. This addresses the shortcomings of traditional repair materials, such as insufficient early guidance or inadequate later-stage support, thus achieving multi-dimensional matching of wound repair needs.

[0011] Preferably, the method for preparing the mildly cross-linked type I collagen includes the following steps: Porcine type I collagen was dissolved in acetic acid solution, cooled to 1-4℃, EDC / NHS solution was added and stirred, pH was adjusted to 4.5-5, and the reaction was carried out for 1.5-2.5 hours. Then, it was dialyzed with acetic acid solution for 45-50 hours to obtain mildly cross-linked type I collagen.

[0012] Preferably, the concentration of EDC is 4-6 mM and the concentration of NHS is 1-3 mM.

[0013] Preferably, the concentration of the acetic acid solution is 0.1M.

[0014] The preparation process of mildly cross-linked type I collagen described in this invention effectively enhances the core efficacy of the final composition, such as skin repair, long-lasting moisturizing, and biocompatibility, by precisely controlling the retention of activity and the degree of cross-linking. This solves the defects of traditional collagen compositions, such as "poor efficacy persistence, high irritation, and weak synergy." Specifically, controlling the low-temperature reaction effectively ensures collagen activity, avoiding composition delamination and efficacy loss due to collagen structure damage; the weakly acidic environment and short reaction time precisely achieve mild cross-linking, forming only a small number of intermolecular cross-linking bonds; the low concentration of EDC / NHS avoids excessive cross-linking, balancing collagen stability and solubility; and the specific concentration ratio of EDC to NHS effectively improves cross-linking efficiency and optimizes collagen structure, thereby working with type III collagen to repair the skin barrier, reduce moisture loss, and fix mussel adhesive protein through the collagen framework, extending its residence time on the skin surface, thus further enhancing the composition's repair persistence.

[0015] Preferably, the humectant comprises sodium hyaluronate; and / or, the preservative comprises at least one of 1,2-pentanediol and 1,2-hexanediol.

[0016] Preferably, the 1,2-pentanediol content in the composition is 1-2% by mass, and the 1,2-hexanediol content in the composition is 0.5-1% by mass.

[0017] Preferably, the type III collagen is recombinant human type III collagen.

[0018] Preferably, the mussel adhesive protein is a recombinant MFP-151 type mussel adhesive protein.

[0019] Preferably, the sodium hyaluronate has a molecular weight of 5.5-6.5 kDa.

[0020] Secondly, the present invention provides the application of the described composition in the preparation of wound repair products.

[0021] Thirdly, the present invention provides a method for preparing the composition, comprising the following steps: S1. Mix mildly cross-linked type I collagen and type III collagen at 1-4℃ and pH 2.8-3.2, and stir at 100-200 rpm for 10-20 min to form the first mixture; S2. After adjusting the pH of the first mixture to 7.2-7.6, add an aqueous solution of mussel adhesive protein containing 80-120mM sodium chloride, and stir at 100-200rpm for 5-15min at 1-4℃ to form the second mixture; S3. Raise the temperature of the second mixture to 35-38℃ and let it stand for 5-7 hours to form a gel. S4. Add humectant and preservative to the gel and stir until homogeneous to obtain the composition.

[0022] This invention employs a stepwise assembly method to mix mildly cross-linked type I and type III collagen in a specific ratio. Under specific pH and temperature conditions, collagen fibers self-assemble to form a preliminary heterogeneous fiber network with type I collagen as the "backbone" and type III collagen as "branches." Subsequently, the catechol groups in a mussel adhesive protein solution containing a low concentration of sodium chloride are slowly oxidized to o-benzoquinone under the action of dissolved oxygen. Under certain ionic strength, some o-benzoquinone can react with nucleophilic groups on the collagen to form covalent cross-links. This results in a three-dimensional network hydrogel with type I collagen as the "skeleton" (providing strength), type III collagen as the "soft network" (providing flexibility), and mussel adhesive protein as the "dynamic cross-linking agent." This structure highly mimics the natural ratio and network structure of type I / III collagen in human dermis, providing the most familiar environment for cell migration and proliferation. The mildly cross-linked type I collagen degrades slightly more slowly in the gel network, serving as a "strong skeleton" in the later stages of repair, providing long-term mechanical support and preventing scar contracture. Type III collagen, due to its high affinity for cells, is preferentially utilized in the early stages of repair, guiding orderly cell migration and proliferation. Mussel adhesive proteins exhibit superior adhesion in moist environments, allowing the gel to firmly adhere to irregular, exudative wounds, forming a physical barrier and maintaining a moist environment. The subsequent addition of humectants and preservatives further enhances the composition's immediate moisturizing and antiseptic effects.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention, for the first time, utilizes a stepwise assembly technique to construct a heterogeneous fiber network from mildly cross-linked type I collagen and type III collagen prepared using a specific process, in a specific ratio. It also innovatively introduces mussel adhesive protein as a dynamic cross-linking agent, enabling self-assembly under physiological conditions to form a three-dimensional hydrogel with a triple structure of "type I collagen skeleton - type III collagen soft network - mussel adhesive protein cross-linking points." This design not only precisely simulates the natural composition and spatial conformation of the dermal extracellular matrix but also achieves sequential functional synergy among the three components—type III collagen guiding cell migration in the early stage, mildly cross-linked type I collagen providing mechanical support in the later stage, and mussel adhesive protein enabling wet adhesion. This breakthrough overcomes the technical bottlenecks of traditional adhesive raw materials in terms of biomimicry, functional synergy, adhesion, and adaptability to wet wounds, providing a superior biomimetic microenvironment for tissue repair and significantly enhancing wound healing efficacy. Attached Figure Description

[0024] Figure 1-3 These are comparison images of some subjects' faces before and after using the composition product described in Example 1. Detailed Implementation

[0025] To better illustrate the objectives, technical solutions, and advantages of this invention, the invention will be further described below with reference to specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

[0026] Unless otherwise specified, all materials and reagents used in the following examples and comparative examples are commercially available and commonly used. Unless otherwise specified, the experimental methods used in the following examples and comparative examples are conventional techniques in the art.

[0027] Example 1 This embodiment provides a composition containing cross-linked collagen, the composition comprising 0.3% mildly cross-linked type I collagen, 0.1% type III collagen, 0.2% mussel adhesive protein, 3% sodium hyaluronate, 2% 1,2-pentanediol, 1% 1,2-hexanediol, and the remainder being water.

[0028] The method for preparing the composition includes the following steps: S1. Dissolve the lyophilized sponge of type I collagen from pigs in 0.1M acetic acid solution, cool it to 4℃, add the predetermined amount of EDC / NHS solution (5mM EDC, 2mM NHS) and stir rapidly. Adjust the pH to 4.7 and react for 2 hours. Then dialyze with 0.1M acetic acid solution for 48 hours to obtain mildly cross-linked type I collagen. S2. Mix mildly cross-linked type I collagen and type III collagen at 4°C and pH=3.0, and stir at 100-200 rpm for 15 min to form the first mixture; S3. After adjusting the pH of the first mixture to 7.4, add an aqueous solution of mussel adhesive protein containing 100mM sodium chloride and stir at 4°C and 150rpm for 10min to form the second mixture. S4. Raise the temperature of the second mixture to 37°C, ventilate in a clean workbench, and let stand for 6 hours to form a gel. S5. Add sodium hyaluronate aqueous solution, 1,2-pentanediol and 1,2-hexanediol to the gel and stir until homogeneous to obtain the composition.

[0029] Example 2 This embodiment provides a composition containing cross-linked collagen, the composition comprising 0.2% mildly cross-linked type I collagen, 0.1% type III collagen, 0.2% mussel adhesive protein, 3% sodium hyaluronate, 2% 1,2-pentanediol, 1% 1,2-hexanediol, and the remainder being water.

[0030] The preparation method of the composition is the same as that in Example 1.

[0031] Example 3 This embodiment provides a composition containing cross-linked collagen, the composition comprising 0.1% mildly cross-linked type I collagen, 0.1% type III collagen, 0.1% mussel adhesive protein, 3% sodium hyaluronate, 2% 1,2-pentanediol, 1% 1,2-hexanediol, and the remainder being water.

[0032] The preparation method of the composition is the same as that in Example 1.

[0033] Example 4 This embodiment provides a composition containing cross-linked collagen, the composition comprising 0.1% mildly cross-linked type I collagen, 0.05% type III collagen, 0.2% mussel adhesive protein, 1% sodium hyaluronate, 1%, 1,2-pentanediol, 0.5% 1,2-hexanediol, and the remainder being water.

[0034] The preparation method of the composition is the same as that in Example 1.

[0035] Comparative Example 1 This comparative example provides a composition comprising 0.3% mildly cross-linked type I collagen, 0.1% type III collagen, 0.2% mussel adhesive protein, 3% sodium hyaluronate, 2% 1,2-pentanediol, 1% 1,2-hexanediol, and the remainder being water.

[0036] The method for preparing the composition includes the following steps: S1. Dissolve the lyophilized sponge of type I collagen from pigs in 0.1M acetic acid solution, cool it to 4℃, add the predetermined amount of EDC / NHS solution (5mM EDC, 2mM NHS), stir rapidly, adjust the pH to 4.7, react for 2 hours, and then dialyze with 0.1M acetic acid solution for 48 hours to obtain mildly cross-linked type I collagen. S2. Mix mildly cross-linked type I collagen, type III collagen, mussel adhesive protein, sodium hyaluronate, 1,2-pentanediol, 1,2-hexanediol and purified water evenly to obtain a composition.

[0037] Comparative Example 2 This comparative example provides a composition comprising 0.3% mildly cross-linked type I collagen, 0.3% mussel adhesive protein, 3% sodium hyaluronate, 2% 1,2-pentanediol, 1% 1,2-hexanediol, and the remainder being water.

[0038] The method for preparing the composition includes the following steps: S1. Dissolve the lyophilized sponge of type I collagen from pigs in 0.1M acetic acid solution, cool it to 4℃, add the predetermined amount of EDC / NHS solution (5mM EDC, 2mM NHS) and stir rapidly. Adjust the pH to 4.7 and react for 2 hours. Then dialyze with 0.1M acetic acid solution for 48 hours to obtain mildly cross-linked type I collagen. S2. After adjusting the pH to 7.4, add an aqueous solution of mussel adhesive protein containing 100mM sodium chloride and stir at 4°C and 100-200rpm for 10min to form the first mixture; S3. Raise the temperature to 37°C, ventilate in a clean bench, and let stand for 6 hours to form a gel. S4. Add sodium hyaluronate aqueous solution, 1,2-pentanediol and 1,2-hexanediol and stir until homogeneous to obtain the composition.

[0039] Comparative Example 3 This comparative example provides a composition comprising 0.3% mildly cross-linked type I collagen, 0.3% type III collagen, 3% sodium hyaluronate, 2% 1,2-pentanediol, 1% 1,2-hexanediol, and the remainder being water.

[0040] The method for preparing the composition includes the following steps: S1. Dissolve the lyophilized sponge of type I collagen from pigs in 0.1M acetic acid solution, cool it to 4℃, add the predetermined amount of EDC / NHS solution (5mM EDC, 2mM NHS) and stir rapidly. Adjust the pH to 4.7 and react for 2 hours. Then dialyze with 0.1M acetic acid solution for 48 hours to obtain mildly cross-linked type I collagen. S2. Mix mildly cross-linked type I collagen and type III collagen at 4°C and pH=3.0, and stir at 100-200 rpm for 15 min to form the first mixture; S3. Raise the temperature to 37℃, slowly adjust the pH to 7.4, stir well, and let stand for 1 hour to form a gel. S4. Add sodium hyaluronate aqueous solution, 1,2-pentanediol and 1,2-hexanediol and stir until homogeneous to obtain the composition.

[0041] Comparative Example 4 This comparative example provides a composition comprising 0.3% type I collagen, 0.1% type III collagen, 0.2% mussel adhesive protein, 3% sodium hyaluronate, 2% 1,2-pentanediol, 1% 1,2-hexanediol, and the remainder being water.

[0042] The method for preparing the composition includes the following steps: S1. Mix porcine type I collagen and type III collagen at 4°C and pH=3.0, and stir at 100-200 rpm for 15 minutes to form the first mixture; S2. After adjusting the pH to 7.4, add an aqueous solution of mussel adhesive protein containing 100mM sodium chloride and stir at 100-200rpm for 10min at 4℃ to form a second mixture; S3. Raise the temperature to 37°C, ventilate in a clean bench, and let stand for 6 hours to form a gel. S4. Add sodium hyaluronate aqueous solution, 1,2-pentanediol and 1,2-hexanediol and stir until homogeneous to obtain the composition.

[0043] Test Example 1 The compositions prepared in Examples 1-4 and Comparative Examples 1-4 were used as test samples, and their efficacy in promoting collagen synthesis was tested using a collagen synthesis amount assay.

[0044] Experimental method: Human primary dermal fibroblasts were expanded and cultured in fibroblast culture medium under the conditions of 5% CO2 and 37℃. When the in vitro cultured fibroblasts reached 80-90% confluence, they were digested with trypsin and seeded into 96-well plates. The cells were cultured in 96-well plates for 48 hours.

[0045] A blank control group, a positive control group, an example group, and a comparative group were set up. The blank control group was serum-free culture medium, the positive control group was 10 ng / mL TGF-β1, and the example and comparative groups were appropriately diluted and added to 200 μL. Each group was replicated in 3 wells. The corresponding culture medium was added to the cell wells of each group, and the cells were incubated in an incubator for 48 h.

[0046] Test method: Cell culture supernatant was used to determine the type I collagen content using the Human Collagen I ELISA kit. The type I collagen content of the example group, comparative group, and positive control group was measured and statistically compared with the blank control group. The relative expression level of type I collagen was calculated using the following formula. All results are expressed as Mean ± SD. The statistical results are shown in Table 1.

[0047] Relative expression level of type I collagen = [(Measured type I collagen content - Added type I collagen content) / Measured type I collagen content in the blank control group] * 100% Table 1 The results in the table above show that the collagen synthesis in the positive control group was significantly higher than that in the blank control group, and the collagen synthesis in the example group was significantly higher than that in the blank control group. Among them, the collagen synthesis in the example 1 group was the highest, indicating that the composition of the present invention can effectively promote the synthesis of type I collagen required for wound repair. The expression levels of Comparative Example 2 (lacking type III collagen) and Comparative Example 3 (lacking mussel adhesive protein) were only 105.8% and 106.4%, respectively, close to the blank control group and far lower than the examples, proving that type III collagen and mussel adhesive protein are the core components that synergistically promote the synthesis of type I collagen with mild cross-linking, and neither can be omitted. Comparative Example 1 did not use a stepwise assembly process, and its expression level was only 109.5%, only slightly higher than the positive control group, indicating that a specific process can ensure that the three components form a stable and effective complex system, directly affecting the final efficacy. Comparative Example 4 used uncross-linked type I collagen, and its expression level was 112.3%, which was higher than some comparative examples, but still far lower than the examples, indicating that mild cross-linking treatment of type I collagen can enhance its interaction with other components and improve the collagen synthesis promotion effect.

[0048] Test Example 2 The composition prepared in Example 1 was used as the test sample, and its anti-enzyme ability was tested by in vitro degradation experiment.

[0049] Methods: Prepare a degradation buffer containing 100 U / mL collagenase (50 mM Tris-HCl, 5 mM CaCl2, 0.15 M NaCl, pH 7.4).

[0050] A blank control group, Example 1 group, and a control group were set up. The blank control group consisted of the sample from Example 1 group in a buffer solution without collagenase, and the control group consisted of uncrosslinked type I collagen lyophilized sponge. Each group had three replicates. 5 mL of collagenase-containing degradation buffer was added to both Example 1 group and the control group, and they were mixed thoroughly. The mixture was then placed on a 37°C constant-temperature shaker with low-speed shaking. The reaction was terminated after 1, 2, 3, and 4 days, respectively. After centrifugation, the supernatant was collected, and the hydroxyproline content was determined by the chloramine-T method. The collagen degradation rate was calculated using the following formula, and the results are shown in Table 2. Collagen degradation rate = [(sample hydroxyproline content - supernatant hydroxyproline content) / sample hydroxyproline content] * 100% Table 2 The results in the table above show that the collagen degradation rate in the Example 1 group was significantly lower than that in the control group under the same enzymatic hydrolysis conditions, indicating that the composition prepared by the present invention can effectively improve the anti-enzyme ability of collagen and prolong the action time.

[0051] Test Example 3 The compositions prepared in Examples 1-4 and Comparative Examples 1-4 were used as test samples, and their repair efficacy was tested using a cell scratch assay.

[0052] Methods: HaCat cells (human immortalized epidermal cells) in logarithmic growth phase were harvested and arranged at a cell density of 2*102. 5 Cells were seeded per well into 24-well culture plates. After culturing in an incubator (37°C, 5% CO2) for 24 h, "damage" was marked by using a 200 μL pipette tip perpendicular to the 24-well plate. The cells were washed three times with PBS solution to remove the marked cells.

[0053] A negative control group, a positive control group, Example 1-4 groups, and Comparative Example 1-4 groups were set up. The negative control group was supplemented with FBS-free medium, the positive control group with medium containing 10% FBS, and the Example and Comparative Example groups were appropriately diluted and added in 100 μL. Each group was divided into three replicates. Cells were incubated in an incubator (37℃, 5% CO2) for 24 h. The migrating cells in each group were photographed using an inverted microscope, and the average scratch area was calculated using Image Pro Plus software. The healing rate was calculated using the following formula, and the results are shown in Table 3. Healing rate = [(Initial scratch area - Current scratch area) / Initial scratch area] * 100% Table 3 The results in the table above show that the cell healing rate of the positive control group was significantly higher than that of the negative control group, and the cell healing rates of the example group and the comparative group were also significantly higher than those of the negative control group. Among them, the cell healing rate of the example group was higher than that of the comparative group 1, which simply mixed the components, and the comparative groups 2 and 3, which lacked one component. This indicates that the composition prepared by the present invention has a special structure, and a better skin repair effect can be achieved through the synergistic effect of the components.

[0054] Test Example 4 The compositions prepared in Examples 1-4 and Comparative Examples 1-4 were used as test samples, and their moisturizing effects were tested using a moisturizing experiment.

[0055] Methods: Participants aged 20-45 years were selected and numbered in groups of five. Prior to the experiment, participants were prohibited from using other similar cosmetics and from engaging in any skincare routine other than routine cleansing. At the start of the experiment, each group of participants used the designated facial cleanser to wash their face and then patted their skin dry with the same facial tissue. Take samples to be tested and select test areas on the face. The test area is 3.0*3.0cm. The product dosage is (0.020±0.001)g. Before the subject uses the test sample (H0), 1 hour after use (H1), 2 hours after use (H2), 4 hours after use (H4), 6 hours after use (H6), and 8 hours after use (H8), the skin stratum corneum moisture content is measured using a skin moisture meter (Corneometer). The results are shown in Table 4.

[0056] Table 4 The results in the table above show that the moisturizing effect of the compositions in the embodiments of the present invention is significantly better than that of the comparative examples. Not only are the initial moisturizing capacity and peak water absorption higher, but the moisturizing effect also decays more slowly over time. In contrast, the moisturizing capacity of the compositions in Comparative Examples 2-3 (lacking type III collagen and mussel adhesive protein, respectively) and Comparative Example 4 (using uncrosslinked type I collagen) is significantly reduced. This demonstrates that type III collagen and mussel adhesive protein are the core components for enhancing the moisturizing effect. Their synergistic effect with the slightly crosslinked type I collagen strengthens the moisturizing capacity, and the slight crosslinking enhances the structural stability of collagen molecules, reducing water evaporation and further improving the longevity of moisturizing. In Comparative Example 1, when the composition was not prepared using the stepwise assembly process of the present invention, its moisturizing performance also decreased, indicating that the specific stepwise assembly process of the present invention can effectively optimize the microscopic three-dimensional structure of the composition, further improving the moisturizing effect.

[0057] Test Example 5 The compositions prepared in Examples 1-4 and Comparative Examples 1-4 were used as test samples, and their wound repair effects were tested using human wound repair experiments.

[0058] Methods: 160 volunteers were selected to participate in a wound repair experiment. Among them, 80 volunteers had undergone phototherapy and minimally invasive surgery, and 80 volunteers had acne. They were randomly divided into 8 groups of 20 people each, and the results of Examples 1-4 and Comparative Examples 1-4 were compared.

[0059] Instructions for use: After cleaning the wound, apply the product to the area that needs repair and gently massage until evenly covered. Use 3 times a day and record the effect after 10 days of use.

[0060] Effect measurement: 1. Significant: 100% wound healing, no allergies, no redness or swelling, and no exudation.

[0061] 2. Effective: 50-80% wound healing rate, no allergies, slight redness and swelling, no exudation.

[0062] 3. Ineffective: Wound healing is less than 50%, with redness, swelling, and exudation. The test results are shown in Table 6. Figure 1-3These are comparison images of some subjects' faces before and after using the composition product described in Example 1.

[0063] Table 6 The results in the table above show that Examples 1-4 have a significantly higher wound repair rate than Comparative Examples 1-4. Among Examples 1-4, the effectiveness of Example 3 is lower than that of the other examples, indicating that the composition of mildly cross-linked type I and type III collagen that simulates the structure ratio of the extracellular matrix has a stronger wound repair ability.

[0064] The results above show that the present invention employs a specific combination and stepwise assembly process of mildly cross-linked type I collagen, type III collagen, and mussel adhesive protein, which can maximize the anti-enzyme, moisturizing, and wound repair capabilities of the composition.

[0065] Finally, it should be noted that the above embodiments are used to illustrate the technical solutions of the present invention and not to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A composition comprising cross-linked collagen, characterized in that, The composition comprises the following components in mass percentage: 0.1-0.3% of slightly cross-linked type I collagen, 0.05-0.1% of type III collagen, 0.1-0.2% of mussel mucin, 1-3% of humectant, 1.5-3% of preservative, and the balance of water.

2. The composition of claim 1, wherein The mass ratio of the slightly cross-linked type I collagen to the type III collagen is (2-3):

1.

3. The composition of claim 2, wherein The mass ratio of the slightly cross-linked type I collagen to the type III collagen is 3:

1.

4. The composition of claim 1, wherein The preparation method of the slightly cross-linked type I collagen comprises the following steps: The pig-derived type I collagen is dissolved with an acetic acid solution, cooled to 1-4℃, and then EDC / NHS solution is added for stirring and mixing, the pH is adjusted to 4.5-5, and the reaction is carried out for 1.5-2.5 hours, and then the slightly cross-linked type I collagen is obtained by dialysis with an acetic acid solution for 45-50 hours.

5. The composition of claim 4, wherein The concentration of the EDC is 4-6mM, and the concentration of the NHS is 1-3mM.

6. The composition of claim 1, wherein The humectant comprises sodium hyaluronate; and / or, the preservative comprises at least one of 1,2-pentanediol and 1,2-hexanediol.

7. Use of the composition according to any one of claims 1-6 in the preparation of a wound repair product.

8. Process for the preparation of a composition according to any one of claims 1 to 6, characterized in that, The method comprises the following steps: S1, mixing the slightly cross-linked type I collagen and the type III collagen at 1-4℃ and pH 2.8-3.2, stirring at 100-200rpm for 10-20min to form a first mixture; S2, adjusting the pH of the first mixture to 7.2-7.6, then adding a mussel mucin aqueous solution containing 80-120mM sodium chloride, stirring at 1-4℃ and 100-200rpm for 5-15min to form a second mixture; S3, raising the temperature of the second mixture to 35-38℃, and standing for 5-7 hours to form a gel; S4, adding the humectant and the preservative to the gel, and stirring uniformly to obtain the composition.

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